A Budding-Defective M2 Mutant Exhibits Reduced Membrane Interaction, Insensitivity To Cholesterol, And Perturbed Interdomain Coupling

Influenza A M2 is a membrane-associated protein with a C-terminal amphipathic helix that plays a cholesterol-dependent role in viral budding. An M2 mutant with alanine substitutions in the C-terminal amphipathic helix is deficient in viral scission. With the goal of providing atomic-level understanding of how the wild-type protein functions, we used a multipronged site-directed spin labeling electron paramagnetic resonance spectroscopy (SDSL-EPR) approach to characterize the conformational properties of the alanine mutant. We spin-labeled sites in the transmembrane (TM) domain and the C-terminal amphipathic helix (AH) of wild-type (WT) and mutant M2, and collected information on line shapes, relaxation rates, membrane topology, and distances within the homotetramer in membranes with and without cholesterol. Our results identify marked differences in the conformation and dynamics between the WT and the alanine mutant. Compared to WT, the dominant population of the mutant AH is more dynamic, shallower in the membrane, and has altered quaternary arrangement of the C-terminal domain. While the AH becomes more dynamic, the dominant population of the TM domain of the mutant is immobilized. The presence of cholesterol changes the conformation and dynamics of the WT protein, while the alanine mutant is insensitive to cholesterol. These findings provide new insight into how M2 may facilitate budding. We propose the AH–membrane interaction modulates the arrangement of the TM helices, effectively stabilizing a conformational state that enables M2 to facilitate viral budding. Antagonizing the properties of the AH that enable interdomain coupling within M2 may therefore present a novel strategy for anti-influenza drug design

Conformational Dynamics of metallo-β-lactamase CcrA during Catalysis Investigated by Using DEER Spectroscopy

Previous crystallographic and mutagenesis studies have implicated the role of a position-conserved hairpin loop in the metallo-β-lactamases in substrate binding and catalysis. In an effort to probe the motion of that loop during catalysis, rapid-freeze-quench double electron–electron resonance (RFQ-DEER) spectroscopy was used to interrogate metallo-β-lactamase CcrA, which had a spin label at position 49 on the loop and spin labels (at positions 82, 126, or 233) 20–35 Å away from residue 49, during catalysis. At 10 ms after mixing, the DEER spectra show distance increases of 7, 10, and 13 Å between the spin label at position 49 and the spin labels at positions 82, 126, and 233, respectively. In contrast to previous hypotheses, these data suggest that the loop moves nearly 10 Å away from the metal center during catalysis and that the loop does not clamp down on the substrate during catalysis. This study demonstrates that loop motion during catalysis can be interrogated on the millisecond time scale

Artificial Aurora Produced by HAARP

We present results from the ionospheric heating experiment conducted at the HighFrequency Active Auroral Research Program (HAARP) facility, Alaska, on 12 March 2013. During theexperiment, HAARP transmitted in the direction of the magnetic zenith X-mode 4.57-MHz wave. Thetransmitted power was modulated with the frequency of 0.9 mHz, and it was pointed on a 20-km spot at thealtitude of 120 km. The heating (1) generates disturbances in the magnetic field detected with the fluxgatemagnetometer on the ground and (2) produces bright luminous spots in the ionosphere, observed with theHAARP telescope. Numerical simulations of the 3-D reduced magnetohydrodynamic (MHD) model revealthat these effects can be related to the magnetic field-aligned currents, excited in the ionosphere bychanging the conductivity in theEregion when the large-scale electric field exists in the heating region

Perseverative cognition and health behaviours: exploring the role of intentions and perceived behavioural control

Objective Worry and rumination (Perseverative Cognition, PC) have been associated with health behaviours, but the underlying mechanisms are unknown. Given the role of physiological experiences on perceived behavioural control (PBC) and emotion regulation on intention-health behaviour relationships, we tested whether: PC prospectively predicts poorer health behaviours; PC moderates the relationship(s) between intentions/PBC and health behaviour, as well as whether the relationship between PC and health behaviour is mediated by intentions and PBC. Methods and Measures In a prospective design, 650 participants (mean age = 38.21 years; 49% female) completed baseline measures of intentions, PBC and PC (worry and rumination) and 590 (mean age = 38.68 years; 50% female) completed follow-up (Time 2) measures of health behaviours (physical activity, sleep, sedentary activity, unhealthy snacking) 1-week later. Results Worry and rumination (at T1) predicted poorer sleep quality. Worry, but not rumination, moderated PBC-physical activity frequency relations. Consistent with mediation, the indirect paths from both worry and rumination, through PBC, to sleep quality and total sleep time were significant. Conclusion PC is associated with poorer sleep quality and PBC can play a mediating role in such relationships. Future research should further consider the role that PBC plays in PC-health behaviour relations

The role of perseverative cognition in the job strain-health outcome relationship

Objective: Job strain has been implicated in a variety of adverse health outcomes, particularly cardiometabolic and inflammatory diseases. However, the mechanisms underlying these effects remain largely unknown. One possibility is that the maladaptive coping response to stress, (perseverative cognition (PC); the cognitive representation of past stressful events (rumination) or feared future events (worry)), either in work or more generally, mediates the relationship between job strain and physical disease. The aim of this study was thus to test the potential role of both general, and work- related PC as a mediating, or potentially moderating, mechanism between job strain and ill- health outcomes. Design & Measures: Using an online cross- sectional design, 650 full- time employees completed measures of job strain, general and work- related PC (rumination & worry) and health outcomes (burnout, somatization, health behaviours & sleep quality). Results: General and work- related worry and rumination significantly mediated, often independently, the relationship between job strain and burnout, somatization, and sleep quality. No significant mediation effects were observed for health behaviours and no type of PC (general or work- related) moderated job strain- health outcome relations. Conclusion: Both general and work- related worry and rumination are likely to play important, and partly independent, roles in understanding the adverse relationships between job strain and various health outcomes

Multiplexed gas spectroscopy using tunable VCSELs

Detection and identification of gas species using tunable laser diode laser absorption spectroscopy has been performed using vertical cavity surface emitting lasers (VCSEL). Two detection methods are compared: direct absorbance and wavelength modulation spectroscopy (WMS). In the first, the output of a DC-based laser is directly monitored to detect for any quench at the targeted specie wavelength. In the latter, the emission wavelength of the laser is modulated by applying a sinusoidal component on the drive current of frequency {omega}, and measuring the harmonics component (2{omega}) of the photo-detected current. This method shows a better sensitivity measured as signal to noise ratio, and is less susceptible to interference effects such as scattering or fouling. Gas detection was initially performed at room temperature and atmospheric conditions using VCSELs of emission wavelength 763 nm for oxygen and 1392 nm for water, scanning over a range of approximately 10 nm, sufficient to cover 5-10 gas specific absorption lines that enable identification and quantization of gas composition. The amplitude and frequency modulation parameters were optimized for each detected gas species, by performing two dimensional sweeps for both tuning current and either amplitude or frequency, respectively. We found that the highest detected signal is observed for a wavelength modulation amplitude equal to the width of the gas absorbance lines, in good agreement with theoretical calculations, and for modulation frequencies below the time response of the lasers (<50KHz). In conclusion, we will discuss limit of detection studies and further implementation and packaging of VCSELs in diode arrays for continuous and simultaneous monitoring of multiple species in gaseous mixtures

A Titanium Nitride Absorber for Controlling Optical Crosstalk in Horn-Coupled Aluminum LEKID Arrays for Millimeter Wavelengths

We discuss the design and measured performance of a titanium nitride (TiN) mesh absorber we are developing for controlling optical crosstalk in horn-coupled lumped-element kinetic inductance detector arrays for millimeter-wavelengths. This absorber was added to the fused silica anti-reflection coating attached to previously-characterized, 20-element prototype arrays of LEKIDs fabricated from thin-film aluminum on silicon substrates. To test the TiN crosstalk absorber, we compared the measured response and noise properties of LEKID arrays with and without the TiN mesh. For this test, the LEKIDs were illuminated with an adjustable, incoherent electronic millimeter-wave source. Our measurements show that the optical crosstalk in the LEKID array with the TiN absorber is reduced by 66\% on average, so the approach is effective and a viable candidate for future kilo-pixel arrays.Comment: 7 pages, 5 figures, accepted for publication in the Journal of Low Temperature Physic

A millimeter-wave kinetic inductance detector camera for long-range imaging through optical obscurants

Millimeter-wave imaging provides a promising option for long-range target detection through optical obscurants such as fog, which often occur in marine environments. Given this motivation, we are currently developing a 150 GHz polarization-sensitive imager using a relatively new type of superconducting pair-breaking detector, the kinetic inductance detector (KID). This imager will be paired with a 1.5 m telescope to obtain an angular resolution of 0.09° over a 3.5° field of view using 3,840 KIDs. We have fully characterized a prototype KID array, which shows excellent performance with noise strongly limited by the irreducible fluctuations from the ambient temperature background. Full-scale KID arrays are now being fabricated and characterized for a planned demonstration in a maritime environment later this year